The gene which encodes (R)-specific carbonyl reductase(rCR) from Candida parapsilosis CCTCC M203011 was cloned.The cloned sequence includes an open reading frame (ORF) consisting of 1011bp, encoding a protein of 336 amino acids, with a molecular weight of 35.9 kD.The nucleotide sequence showed 99% similarity to those of the other members of the alcohol dehydrogenase superfamily.A recombinant Escherichia coli JM109 strain harboring the expression plasmid, produced (R)-1-phenyl-1,2-ethanediol(100%e.e,80.4%yield) from ?-hydroxyacetophenone without any additive to regenerate NAD~ + from NADH.

It is now well established that inflammation plays an important role in the development of numerous chronic metabolic diseases including insulin resistance (IR) and type 2 diabetes (T2DM). Skeletal muscle is responsible for 75% of total insulin-dependent glucose uptake; consequently, skeletal muscle IR is considered to be the primary defect of systemic IR development. Our pre- vious study has shown that rutaecarpine (Rut) can benefit blood lipid profile, mitigate inflammation, and improve kidney, liver, pan- creas pathology status of T2DM rats. However, the effects of Rut on inflammatory cytokines in the development of IR-skeletal muscle cells have not been studied. Thus, our objective was to investigate effects of Rut on inflammatory cytokines interleukiri (IL)-1, IL-6 and tumor necrosis factor (TNF)-α in insulin resistant primary skeletal muscle cells (IR-PSMC). Primary cultures of skeletal muscle cells were prepared from 5 neonate SD rats, and the primary rat skeletal muscle cells were identified by cell morphology, effect of ru- taecarpine on cell proliferation by MTT assay. IR-PSMC cells were induced by palmitic acid (PA), the glucose concentration was measured by glucose oxidase and peroxidase (GOD-POD) method. The effects of Rut on inflammatory cytokines IL-1, IL-6 and TNF-α in IR-PSMC cells were tested by enzyme-linked immunosorbent assay (ELISA) kit. The results show that the primary skeletal muscle cells from neonatal rat cultured for 2-4 days, parallel alignment regularly, and cultured for 7 days, cells fused and myotube formed. It was shown that Rut in concentration 0-180. 0 μmol x L(-1) possessed no cytotoxic effect towards cultured primary skeletal muscle cells. However, after 24 h exposure to 0.6 mmol x L(-1) PA, primary skeletal muscle cells were able to induce a state of insulin resistance. The results obtained indicated significant decrease (P < 0.05 to P < 0.001) IL-1, IL-6 and TNF-α production by cultured IR-PSMC cells when incubating 24 hours with Rut, beginning from 20 to 180.0 μmol x L(-1). IL-1, IL-6 and TNF-α in the Rut treated groups were dose-dependently decreased compared with that in the IR-PSMC control group. Our results demonstrated that the Rut promoted glucose consumption and improved insulin resistance possibly through suppression of inflammatory cytokines in the IR-PSMC cells.
Animals ; Cell Proliferation ; drug effects ; Cytokines ; metabolism ; Female ; Glucose ; metabolism ; Indole Alkaloids ; pharmacology ; Inflammation ; metabolism ; Insulin Resistance ; Male ; Muscle, Skeletal ; cytology ; drug effects ; metabolism ; Quinazolines ; pharmacology ; Rats

OBJECTIVETo investigate the efficacy and safety of half-dose Zenapax for prevention of acute rejection after renal transplantation.

METHODSAccording to the immunosuppressive regimen and renal function after transplantation, patients were divided into 4 groups, namely groups A, B, C, and D of 90, 73, 11 and 13 patients, respectively. Blood creatinine measured 1 week after operation was <176.6 micromol/L in groups A and B, and was >353 micromol/L in groups C and D. Patients in groups A and C were given 25 mg Zenapax (0.5 mg/kg) and MMF 0.75 g before operation, and those in groups B and D had only MMF of 0.75 g. All patients were given Pred, CsA and MMF after operation, and the rejection episodes, the time of acute rejection onset, the rate of rejection reversal and complications were analyzed in the time period of 6 months after operation.

RESULTSAfter the operation, 13 patients (14.4%) developed acute rejection in group A, 18 (24.6%) in group B, 6 (54.5%) in group C and 7 (53.8%) in group D (P<0.01). The incidence of acute rejection in group B was significantly lower than that in groups C and D groups (P<0.01), and the latter two groups had similar incidence. The time of acute rejection onset ranged from 3 to 9 days postoperatively (mean 6.2-/+3.2 days) in group A, significantly delayed as compared with that in group B (range 2-8 days, mean 4.7-/+3.1 days), group C (range 2-7 days, mean 4.3-/+4.2 days) and group D group (range 2-9 days, mean 3.9-/+3.5 days), but the time was similar between groups B, C, and D (P>0.05). All acute rejection cases in group A was reversed, and the rate of reversal was 88.9% (16/18) in group B, 83.3% in group C, and 71.4% in group D. No significant differences were noted in such complications as infection, vascular injuries or gastrointestinal reactions between the 4 groups (P>0.05).

CONCLUSIONZenapax at the dose of 25 mg can safely decrease the risk of acute rejection in patients with good postoperative renal function recovery, but dose not seem effective in patients with delayed graft function recovery.

Acute Disease ; Adolescent ; Adult ; Antibodies, Monoclonal ; administration & dosage ; Antibodies, Monoclonal, Humanized ; Creatinine ; blood ; Female ; Follow-Up Studies ; Graft Rejection ; etiology ; prevention & control ; Humans ; Immunoglobulin G ; administration & dosage ; Immunosuppressive Agents ; administration & dosage ; Kidney Transplantation ; adverse effects ; methods ; Male ; Middle Aged ; Postoperative Complications ; etiology ; prevention & control ; Treatment Outcome

Objective To evaluate the outcome of health education program on drowning prevention among primary and secondary school children in rural areas. Methods A township was selected and all the students from grade 3 to 5, grade 7 to 8, and grade 10 to 11 were selected to take part in the program. Twelve intervention measures on natural water safety and drowning prevention were carried out for one year. Information was collected using the same questionnaire before and after the intervention program. Results One year after the intervention was carried out, children's knowledge on drowning prevention improved significantly (13.21, 95% CI: 12.51-13.90) , and a positive effect was also noticed among boys (12.77, 95%CI: 11.77-13.77), girls (13.80, 95%CI: 12.82-14.78),and among primary school children (15.51,95%CI: 14.30-16.72), senior high school children (10.78,95%CI: 9.50-12.05) and junior high school children (12.77,95%CI: 11.84-13.71). Overall rates on risk behaviors dropped from 41.4% to 32.2% (by 22.2%) including 15.6% for boys, 35.2% for girls and 13.8%, 29.3%, 26.3% for primary school children, senior high school children, junior high school children, respectively. The incidence rates for non-fatal drowning decreased by 58.9% (from 5.6% to 2.3%). The person-times for treatment on sight, in emergency settings, in outpatient clinic or in the hospitals had a reduction from 399, 78, 36 to 175, 32, 14, respectively. Conclusion Health education program could improve children's perception on water safety, and reduce their risk behaviors as well as on the incidence of non-fatal drowning in the rural areas.

In order to develop a rapid detection kit for novel avian influenza virus (AIV) subtype H7N9, two sets of specific primers and probes were designed based on the nucleotide sequences of hemagglutinin antigen (HA) and neuraminidase antigen (NA) of novel H7N9 virus (2013) available in GenBank to establish the method of TaqMan probe-based multiplex real-time RT-PCR for rapid detection of AIV subtype H7N9. The primer and probe of HA were for all H7 subtype AIVs, while the primer and probe of NA were only for novel N9 subtype AIVs. The results showed that this method had high sensitivity and specificity. This method was applicable to the testing of positive standard sample with a minimum concentration of 10 copies/microL; it not only distinguished H7 subtype from H1, H3, H5, H6, and H9 subtypes, but also distinguished novel N9 subtype from traditional N9 subtype. A total of 2700 samples from Zhuhai, China were tested by this method, and the results were as expected. For the advantages of sensitivity and specificity, the method holds promise for wide application.
Animals ; Birds ; virology ; Influenza A Virus, H7N9 Subtype ; genetics ; isolation & purification ; physiology ; Influenza in Birds ; prevention & control ; virology ; Real-Time Polymerase Chain Reaction ; methods ; Species Specificity ; Taq Polymerase ; metabolism ; Time Factors

OBJECTIVETo investigate the serum levels of cholesterol, triglyceride and high density lipoprotein cholesterol in children aged 3-14 years and its influencing factors.

METHODSThe cross-sectional survey study population was a representative sample from Guangdong province obtained by multi-stage randomized cluster sampling. Serum lipids in 6188 children aged > or = 3 years were assayed using automatic biochemical instrument. The data of social and demographic status were collected by face-to-face interview, and height and weight were obtained by physical examination.

RESULTSThe age-standardized and region-weighted means of serum total cholesterol (TC), triglyceride (TG), and high density lipoprotein cholesterol (HDL-C) were 0.80 mmol/L, 3.50 mmol/L and 1.28 mmol/L, respectively. For the mean of TG, there was no difference between metropolitan and middle city, nor between rich county and poor county. For TC, it was the highest in metropolitan, and there was no difference between rich and poor county. For HDL-C, the difference existed between every two regions. The age-standardized and region-weighted prevalence of high TG, high TC and low HDL-C were 2.2%, 2.1% and 8.0%, respectively. Metropolitan, rich county, low weight and age between 7.0-9.9 years are protecting factors for high TG, and the number of family between 3-4, age between 7.0-9.9, metropolitan, middle city and poor county are risk factors for high TC. Male, family income per year between 800-9999 RMB, middle city, rich county are protecting factors for low HDL-C.

CONCLUSIONThe prevalence of abnormal serum lipid was still low compared with other regions in China. The region, number of family member, age and sex may be the important factors influencing on serum lipid levels.

Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Cross-Sectional Studies ; Dyslipidemias ; epidemiology ; Female ; Humans ; Male ; Risk Factors ; Triglycerides ; blood

OBJECTIVETo describe the epidemiological characteristics of overweight and obesity among people aged 18 and over in Guangdong province in 2002, and to identify the populations and regions under high risk.

METHODSCross-sectional survey was used through sampling on multi-stage randomized clusters. Data of socialeconomic status were collected by face-to-face interview. Data on weight and height was obtained through physical check-ups.

RESULTSA sample size of 15 130 people and the mean body mass index (BMI) was 22.03 +/- 3.38 with no significant difference between males and females. However the significant difference was seen between cities and rural areas. The crude prevalence rate of overweight and obesity were 16.8% and 1.8%, and the age-adjusted rate were 15.0% and 1.7%, respectively. The crude rate of overweight in cities (24.8%) and males (17.5%) were higher than that in rural areas (9.4%) and females (16.2%). The crude rate of obesity in cities was seen higher than that in the rural areas, but not significantly different between females and males. The results of logistic regression analysis showed that the major risk factors influencing overweight would include household income, age, gender, smoking habits, physical exercises and location of residence.

CONCLUSIONAbout one sixth of the citizens in Guangdong province were considered to be overweighted and obesive had become an important public health problem. Integrated measures must be taken for prevention and control.

Adolescent ; Adult ; Body Mass Index ; China ; epidemiology ; Cross-Sectional Studies ; Diet ; Exercise ; Female ; Humans ; Male ; Obesity ; epidemiology ; Overweight ; Prevalence ; Risk Factors ; Sampling Studies ; Socioeconomic Factors ; Surveys and Questionnaires

OBJECTIVETo investigate the optimal conditions for establishing insulin-resistant 3T3-L1 adipocytes.

METHOSDexamethason (DEX), 3-isobutyl-methylxanthine (IBMX) and different concentrations of insulin (10(-8), 10(-7), and 10(-6) mol·L(-1)) were used to induce 3T3-L1 preadipocytes into mature adipocytes identified by oil red O staining. We established insulin- resistant 3T3-L1 adipocytes cell model (IR-3T3-L1) by exposing the cells to 1µmol·L(-1) DEX, and the changes of glucose concen- tration in the cell culture were determined by glucose oxidase-peroxidase (GOD-POD) assay.

RESULTSTreatment of 3T3-L1 cells with DEX, IBMX and 10(-6) mol·L(-1)) insulin for 9 days resulted in the differentiation of >90% of the cells into mature adipocytes. IR-3T3-L1 cells cultured for 96 h in the culture media containing 1 µmol·L(-1) DEX showed significantly increased glucose consumption (P=0.0003) as compared with the control group at 36 h (P<0.001).

CONCLUSION3T3-L1 cells can be induced into mature adipocytes by exposure to 1 µmol·L(-1) DEX, 0.5 mmol·L(-1) IBMX and 10(-6) mol·L(-1)) insulin. A 96 h exposure to 1 µmol·L(-1) DEX can induce 3T3-L1 adipocytes to acquire insulin resistance that can be maintained for 36 h.

1-Methyl-3-isobutylxanthine ; chemistry ; 3T3-L1 Cells ; Adipocytes ; cytology ; Animals ; Cell Differentiation ; Culture Media ; chemistry ; Dexamethasone ; chemistry ; Glucose ; chemistry ; Insulin ; chemistry ; Insulin Resistance ; Mice

In order to study the proliferation inhibition effect of H5N1 subtype avian influenza virus (AIV) with small interfere RNA (siRNA), a total of 4 siRNAs were designed in accordance with the NP and PA genes of H5N1 subtype AIV, the siRNAs were then transfected to chicken embryo fibroblast(CEF), CEF was infected with H5N1 subtype AIV after 6 hrs. Virus titer of cell supernatant was tested at 16-56hrs post infection, and pathological changes of the cells was observed; mRNA levels of NP, PA, HA and p13-actin gene were tested at 36hrs post infection. The results showed that these 4 siRNAs could inhibit the prolif-eration of H5N1 subtype AIV in CEF in varying degrees, and one siRNA targeting PA was best per-formed. The experimental results also showed that the inhibition effect was decreased with the time prolonged. This research provides a basis for further studying RNAi on AIV prevention and control.
Actins ; genetics ; Animals ; Chick Embryo ; DNA Primers ; genetics ; Fibroblasts ; virology ; Hemagglutination ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Hemagglutinins ; genetics ; Humans ; Influenza A Virus, H5N1 Subtype ; genetics ; growth & development ; physiology ; RNA Interference ; RNA Replicase ; genetics ; RNA, Small Interfering ; chemical synthesis ; genetics ; RNA-Binding Proteins ; genetics ; Real-Time Polymerase Chain Reaction ; Specific Pathogen-Free Organisms ; Transfection ; Viral Core Proteins ; genetics ; Viral Proteins ; genetics ; Virus Replication

BACKGROUNDAccurate assessment of intra- as well as extra-cardiac malformations and radiation dosage concerns are especially crucial to infants and children with interrupted aortic arch (IAA). The purpose of this study is to investigate the value of prospective electrocardiogram (ECG)-triggered dual-source computed tomography (DSCT) angiography with low-dosage techniques in the diagnosis of IAA.

METHODSThirteen patients with suspected IAA underwent prospective ECG-triggered DSCT scan and transthoracic echocardiography (TTE). Surgery was performed on all the patients. A five-point scale was used to assess image quality. The diagnostic accuracy of DSCT angiography and TTE was compared with the surgical findings as the reference standard. A nonparametric Chi-square test was used for comparative analysis. P <0.05 was considered as a significant difference. The mean effective radiation dose (ED) was calculated.

RESULTSDiagnostic DSCT images were obtained for all the patients. Thirteen IAA cases with 60 separate cardiovascular anomalies were confirmed by surgical findings. The diagnostic accuracy of TTE and DSCT for total cardiovascular malformations was 93.7% and 97.9% (P > 0.05), and that for extra-cardiac vascular malformations was 92.3% and 99.0% (P < 0.05), respectively. The mean score of image quality was 3.77 ± 0.83. The mean ED was 0.30 ± 0.04 mSv (range from 0.23 mSv to 0.39 mSv).

CONCLUSIONSIn infants and children with IAA, prospective ECG-triggered DSCT with low radiation exposure and high diagnostic efficiency has higher accuracy compared to TTE in detection of extra-cardiac vascular anomalies.

Aorta, Thoracic ; diagnostic imaging ; pathology ; Coronary Angiography ; methods ; Electrocardiography ; methods ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Radiation Dosage